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Expressions for Multilinear Combined pH/Organic Solvent Elution of Ionizable Analytes in Reversed-Phase HPLC

Zisi, Ch., Fasoula, S., Pappa-Louisi, A., Nikitas, P.
Analytical chemistry 2013 v.85 no.20 pp. 9514-9521
amino acids, data collection, pH, reversed-phase high performance liquid chromatography, reversed-phase liquid chromatography, solutes, solvents, temporal variation
Expressions for the retention time of ionogenic analytes eluted under multilinear double pH/solvent-gradients in reversed-phase liquid chromatography are developed by dividing each gradient profile into a finite number of subportions, where the solute retention factors or their logarithms vary linearly with time. To test the theory, two series of experimental gradient retention data of amino acid OPA derivatives were analyzed: The first one was a monolinear or bilinear pH-gradient data set obtained in eluents with different but constant organic modifier contents, whereas the second data set comprised retention data of combined pH/organic solvent-gradients, where the organic content was changed linearly with time but the variation of pH exhibited a curved form approximated by five linear subportions. It was found that the derived expressions describe these experimental retention data with high accuracy, since under double pH/solvent-gradients the overall errors in the fitted and predicted retention times were 1.9% and 1.7%, respectively, whereas under simple pH-gradients these errors were 0.9% and 2%, respectively.